An improved, rapid competitive ELISA using a novel conserved 3B epitope for the detection of serum antibodies to foot-and-mouth disease virus

Authors: CHUNG, CHUNGWON - Us Deparment Of Homeland Security; CLAVIJO, ALFONSO - Institute For Infectious Animal Diseases; BOUNPHENG, MANGKEY - Institute For Infectious Animal Diseases; UDDOWLA, SABENA - Oak Ridge Institute For Science And Education (ORISE); SAYED, ABU - Animal And Plant Health Inspection Service (APHIS); DANCHO, BROOKE - Animal And Plant Health Inspection Service (APHIS); OLESEN, IAN - Animal And Plant Health Inspection Service (APHIS); PACHECO, JUAN - Oak Ridge Institute For Science And Education (ORISE); KAMICKER, BARBARA - Leidos; BRAKE, DAVID - Bioquest Associates, Llc; MUDIYANSELAGE, CAREY - Veterinary Medical Research & Development, Inc (VMRD); LEE, STEPHEN - University Of Idaho; RAI, DEVENDRA - Former ARS Employee; Rieder, Aida - Elizabeth

Submitted to: Journal of Veterinary Diagnostic Investigation
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/26/2018
Publication Date: 6/19/2018
Citation: Chung, C.J., Clavijo, A., Bounpheng, M., Uddowla, S., Sayed, A., Dancho, B., Olesen, I.C., Pacheco, J., Kamicker, B.J., Brake, D.A., Bandaranayaka-Mudiyanselage, C., Lee, S.S., Rai, D.K., Rieder, A.E. 2018. An improved, rapid competitive ELISA using a novel conserved 3B epitope for the detection of serum antibodies to foot-and-mouth disease virus. Journal of Veterinary Diagnostic Investigation. 30(5):699-707. https://doi.org/10.1177/1040638718779641.
DOI: https://doi.org/10.1177/1040638718779641

Interpretive Summary: Foot-and-mouth disease (FMD) is a devastating vesicular disease caused by a viruys (FMDV). Its introduction into US could be devastating to livestock, agriculture and other related industries. Readily available diagnostics are important to prevent and control FMD. Serological diagnostic tests used in the US have to be imported from foreign manufacturers, have a high cost and require overnight incubation. ARS scientists were part of a consortium of government, academic institutions and industry to develop and validate a competitive ELISA (cELISA) that detects antibodies only produced in FMDV infected animals. The test targets a unique viral protein (called 3ABC) using a specific monoclonal blocking antibody and a non-infectious antigen. The test has been optimized to provide results after only 90 min serum incubation. The performance data demonstrated that this rapid (3 h), select agent-free assay can be a pivotal tool for FMD surveillance and emergency response. USDA review of pre-licensing serials and validation data resulted in a U.S. Veterinary Biological Product License.

Technical Abstract: The highly contagious foot-and-mouth disease virus (FMDV) afflicts cloven-hoofed livestock and wildlife resulting in heavy economic losses due to loss of trade and recovery from disease. We developed a rapid, sensitive, and specific competitive ELISA to detect serum antibodies to FMDV. This new cELISA utilized a FMDV 3B epitope specific monoclonal blocking antibody and a mutated FMDV 3ABC coating protein at optimized format variables including a 90 min serum incubation. Sera from eight species of animals positive for antibodies to non-FMD vesicular diseases were negative in this assay, demonstrating analytical specificity. The 3B cELISA detected FMDV antibodies in sera from animals infected with one of four tested serotypes by 7-11 d post-infection, demonstrating analytical sensitivity. The diagnostic sensitivity was 100% against sera from cattle infected with one of seven FMDV serotypes, emphasizing serotype-agnostic results. The diagnostic specificities with U.S. cattle and pig sera were 99.4 and 100%, respectively. High repeatability and reproducibility in 3B cELISA results were demonstrated from 3.1% coefficient of variation in percent inhibition data and 100% agreement in positive or negative decision, with no difference between bench and biosafety cabinet operation. DIVA capability was confirmed by negative results in sera from vaccinated, uninfected cattle, pigs, and sheep. This rapid (<3 h), select agent-free assay with high sensitivity, specificity and DIVA feasibility should be a useful tool in surveillance and emergency preparedness in FMDV control programs.